Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity
In recent years, researchers have leveraged the ubiquitin-proteasome system (UPS) to induce selective degradation of proteins by E3 ubiquitin ligases, which has great potential as novel therapeutics for human diseases, including cancer and neurodegenerative disorders. However, despite extensive effo...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/4/479 |
| _version_ | 1797436733114023936 |
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| author | Bayonle Aminu Julia Fux Evan Mallette Nathaniel Petersen Wei Zhang |
| author_facet | Bayonle Aminu Julia Fux Evan Mallette Nathaniel Petersen Wei Zhang |
| author_sort | Bayonle Aminu |
| collection | DOAJ |
| description | In recent years, researchers have leveraged the ubiquitin-proteasome system (UPS) to induce selective degradation of proteins by E3 ubiquitin ligases, which has great potential as novel therapeutics for human diseases, including cancer and neurodegenerative disorders. However, despite extensive efforts, only a handful of ~600 human E3 ligases were utilized, and numerous protein–protein interaction surfaces on E3 ligases were not explored. To tackle these problems, we leveraged a structure-based protein engineering technology to develop a multi-domain fusion protein bringing functional E3 ligases to the proximity of a target protein to trigger its proteasomal degradation, which we termed Ubiquitin Variant Induced Proximity (UbVIP). We first generated non-inhibitory synthetic UbV binders for a selected group of human E3 ligases. With these UbVs employed as E3 ligase engagers, we designed a library of UbVIPs targeting a DNA damage response protein 53BP1. We observed that two UbVIPs recruiting RFWD3 and NEDD4L could effectively induce proteasome degradation of 53BP1 in human cell lines. This provides a proof-of-principle that UbVs can act as a means of targeted degradation for nucleus-localized proteins. Our work demonstrated that UbV technology is suitable to develop protein-based molecules for targeted degradation and can help identify novel E3 ligases for future therapeutic development. |
| first_indexed | 2024-03-09T11:06:48Z |
| format | Article |
| id | doaj.art-23f11e7cfc334a04892f52598fcca073 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T11:06:48Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-23f11e7cfc334a04892f52598fcca0732023-12-01T00:55:07ZengMDPI AGBiomolecules2218-273X2022-03-0112447910.3390/biom12040479Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced ProximityBayonle Aminu0Julia Fux1Evan Mallette2Nathaniel Petersen3Wei Zhang4Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, CanadaDepartment of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, CanadaDepartment of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, CanadaDepartment of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, CanadaDepartment of Molecular and Cellular Biology, College of Biological Science, University of Guelph, 50 Stone Rd E, Guelph, ON N1G 2W1, CanadaIn recent years, researchers have leveraged the ubiquitin-proteasome system (UPS) to induce selective degradation of proteins by E3 ubiquitin ligases, which has great potential as novel therapeutics for human diseases, including cancer and neurodegenerative disorders. However, despite extensive efforts, only a handful of ~600 human E3 ligases were utilized, and numerous protein–protein interaction surfaces on E3 ligases were not explored. To tackle these problems, we leveraged a structure-based protein engineering technology to develop a multi-domain fusion protein bringing functional E3 ligases to the proximity of a target protein to trigger its proteasomal degradation, which we termed Ubiquitin Variant Induced Proximity (UbVIP). We first generated non-inhibitory synthetic UbV binders for a selected group of human E3 ligases. With these UbVs employed as E3 ligase engagers, we designed a library of UbVIPs targeting a DNA damage response protein 53BP1. We observed that two UbVIPs recruiting RFWD3 and NEDD4L could effectively induce proteasome degradation of 53BP1 in human cell lines. This provides a proof-of-principle that UbVs can act as a means of targeted degradation for nucleus-localized proteins. Our work demonstrated that UbV technology is suitable to develop protein-based molecules for targeted degradation and can help identify novel E3 ligases for future therapeutic development.https://www.mdpi.com/2218-273X/12/4/479Ubiquitin Varianttargeted protein degradationPROTACE3 ligase53BP1NEDD4L |
| spellingShingle | Bayonle Aminu Julia Fux Evan Mallette Nathaniel Petersen Wei Zhang Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity Biomolecules Ubiquitin Variant targeted protein degradation PROTAC E3 ligase 53BP1 NEDD4L |
| title | Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity |
| title_full | Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity |
| title_fullStr | Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity |
| title_full_unstemmed | Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity |
| title_short | Targeted Degradation of 53BP1 Using Ubiquitin Variant Induced Proximity |
| title_sort | targeted degradation of 53bp1 using ubiquitin variant induced proximity |
| topic | Ubiquitin Variant targeted protein degradation PROTAC E3 ligase 53BP1 NEDD4L |
| url | https://www.mdpi.com/2218-273X/12/4/479 |
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